Compositions and methods for treating and preventing crimean congo hemorrhagic fever virus

Abstract

Disclosed herein are immunogenic compositions or product combinations of engineered CCHFV nucleic acids, genes, peptides, or proteins that can be used to elicit an immune response against a CCHFV infection. Also disclosed are methods of using the immunogenic compositions or product combinations in subjects to generate immune responses against CCHFV by administering the compositions or combinations with a nucleic acid prime and polypeptide boost approach.

Description

COMPOSITIONS AND METHODS FOR TREAT ING AND PREVENTINGCRIMEAN CONGO HEMORRHAGIC FEVER VIRUSINCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Ser. No. 63 / 488635, filed March 6, 2023, which is hereby expressly incorporated by reference in its entirety.REFERENCE TO SEQUENCE LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided in a file entitled SeqListmgSVF009WO.xml, which was created on February 26, 2023 and is 79,567 bytes in size. The information in the electronic Sequence Listing is hereby expressly incorporated by reference in its entirety.BACKGROUND OF THE INVENTIONField of the Invention

[0003] Aspects of the present disclosure relate generally to immunogenic compositions or product combinations of engineered Crimean Congo Hemorrhagic fever virus (CCHFV) nucleic acids, genes, peptides, or proteins that can be used to elicit an immune response against CCHFV. This immune response includes activation of cytotoxic immune cells and immune cells that produce neutralizing antibodies against CCHFV. The disclosure also relates generally to methods of using the immunogenic compositions or product combinations in subjects to generate immune responses against CCHFV by administering the compositions or combinations with a homologous or heterologous nucleic acid and / or polypeptide prime and nucleic acid and / or polypeptide boost approach.Description of the Related Art

[0004] The Crimean Congo Hemorrhagic Fever Virus (CCHFV) is an enveloped negative stranded RNA virus with a helical capsid symmetry belonging to the Bunyaviruses that causes a severe and often lethal hemorrhagic disease (1). CCHFV is transmitted by ticksand its epidemiology has expanded from Africa and the Middle-East to Europe and Asia, possibly due to climate changes. There is today no globally available vaccine.

[0005] It has been shown that a DN A vaccine containing two plasmids containing the M-segment containing the GnGc envelope protein and the N gene can protect mice and non-human primates (NHPs) against severe disease (2). In addition, the individual GnGc and N genes have been shown to protect mice against lethal disease either as DNA or mRNA vaccines (3). In NHPs, DNA vaccination with the individual M and N genes were found to reduce viral replication, whereas vaccination with a combination of both genes were found to protect better against disease markers after only two vaccinations (4). Thus, the M and N gene segments are strongly associated with immune responses that protect against disease.

[0006] The mechanism of protection against disease in Crimean Congo Hemorrhagic Fever (CCHF) is not fully understood. It has been shown that immune responses to the N protein alone can protect against lethal disease in mice and can reduce viral replication in NHPs (4). In two vaccine studies, NHPs failed to show a strong induction of neutralizing antibodies despite protection against disease (2, 4). In addition, immune responses against the GP38 protein, encoded by the M segment, has shown protective properties in the absence of neutralizing antibodies (5,6). The need for immunogenic compositions that prevent, inhibit, ameliorate or treat CCHFV infection, or a sequela associated therewith is manifest.SUMMARY

[0007] Disclosed herein are several therapeutic agents useful in immunogenic compositions that prevent, inhibit, ameliorate or treat CCHFV infection, or a sequela associated therewith, as well as methods of using these immunogenic compositions in subjects, such as humans infected with CCHFV or at risk of being infected by CCHFV. .Also described herein are heterologous prime-boost immunization approaches, which utilize a nucleic acid (DNA or RNA) prime and a polypeptide boost administration schedule.

[0008] In some embodiments, the immunogenic compositions or product combinations described herein comprise non-naturally occurring composite nucleic acids and / or polypeptides. In some embodiments, the nucleic acids are DNA or RNA. In some embodiments, the immunogenic compositions or product combinations are administered to an animal, such as a mammal, mouse, rabbit, cat, dog, or human, to induce an immunogenicresponse against CCHFV. In some embodiments, the immunogenic response comprises, consists essentially of, or consists of formation of active immune cells, such as cytotoxic T cells or immune cells that produce inactivating antibodies against the CCHFV, or any antigen, polypeptide, protein, nucleic acid, or genome component of CCHV. In some embodiments, the immunogenic compositions or product combinations are administered to subjects that are at risk of contracting CCHFV or are not currently infected with CCHFV. In some embodiments, the immunogenic compositions or product combinations disclosed herein provide lasting immunogenic protection against a CCHFV infection.

[0009] Some alternatives described herein concern nucleic acids comprising, consisting essentially of, or consisting of at least one CCHFV nucleic acid component. In some alternatives, the at least one CCHFV nucleic acid component comprises, consists essentially of, or consists of an S-segment protein sequence, and / or M-segment protein sequence. In some alternatives, the at least one CCHFV nucleic acid component contemplated for inclusion in the compositions and the methods described herein are human codon optimized sequences of the aforementioned wild-type sequences. In some alternatives, for example, the nucleic acids share at least 50%, 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ I® NO: 18, 21, 24, 27, 30, or 33 or an amount of sequence identity' to any one or more of SEQ ID NO: 18, 21, 24, 27, 30, or 33 that is within a range defined by any two of the aforementioned percentages, wherein said nucleic acids encode an immunogenic or antigenic polypeptide, epitope, or fragment of CCHFV. In some alternatives, the nucleic acids referenced above are used for the prevention, treatment, amelioration, or inhibition of a CCHFV infection or sequela associated therewith in a subject, such as a mammal, preferably a human. Accordingly, some alternatives include the use of a nucleic acid having at least 50%, 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to any one or more of SEQ ID NO: 18, 21, 24, 27, 30, or 33 or an amount of sequence identity to any one or more of SEQ ID NO: 18, 21, 24, 27, 30, or 33 that is within a range defined by any two of the aforementioned percentages, wherein said nucleic acids encode an immunogenic or antigenic polypeptide, epitope, or fragment of CCHFV, as a medicament, such as for the prevention, treatment, amelioration, or inhibition of a CCHFV infection or a sequela associated therewith, in a subject, such as a mammal, preferably a human, which may,optionally, be selected or identified to receive a medicament for the prevention, treatment, amelioration, or inhibition of a CCHFV infection or a sequela associated therewith. Such subjects can be selected or identified by clinical evaluation or diagnostic evaluation or both.

[0010] Some alternatives provided herein concern polypeptides comprising, consisting essentially of, or consisting of at least one CCHFV polypeptide component. In some alternatives, the at least one CCHFV polypeptide component comprises, consists essentially of, or consists of an S-segment protein sequence and / or M-segment protein sequence. In some embodiments, the polypeptides, which may be provided in a composition or method described herein share at least 50%, 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to any one or more of SEQ ID NO: 19, 22, 25, 28, 31, or 34 or an amount of sequence identity to any one or more of SEQ ID NO: 19, 22, 25, 28, 31, or 34 that is within a range defined by any two of the aforementioned percentages, wherein said polypeptides encode an immunogenic or antigenic portion, epitope, or fragment of CCHFV. In some alternatives, the polypeptides are used as a medicament, such as for the prevention, treatment, amelioration, or inhibition of CCHFV or a sequela associated therewith in a subject such as a mammal, preferably a human, which may, optionally, be selected or identified to receive a medicament for the prevention, treatment, amelioration, or inhibition of a CCHFV infection or a sequela associated therewith. Such subjects can be selected or identified by clinical evaluation or diagnostic evaluation or both. In some embodiments, the polypeptides are translated from the wild-type or codon optimized sequences referenced above. In some embodiments, the polypeptides are recombmantly expressed. In some embodiments, the polypeptides are recombmantly expressed in a mammalian, bacterial, yeast, insect, or cell-free system. Accordingly, some alternatives include the use of a polypeptide having at least 50%, 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to any one or more of SEQ ID NO: 19, 22, 25, 28, 31, or 34 or an amount of sequence identity to any one or more of SEQ ID NO: 19, 22, 25, 28, 31, or 34 that is within a range defined by any two of the aforementioned percentages, wherein said nucleic acids encode an immunogenic or antigenic portion, epitope, or fragment of CCHFV as a medicament, such as for the prevention, treatment, amelioration, or inhibition of a CCHFV infection or a sequela associated therewith in a subject, such as a mammal, preferably a human, which may,optionally, be selected or identified to receive a medicament for the prevention, treatment, amelioration, or inhibition of a CCHFV infection or a sequela associated therewith.

[0011] In some alternatives, the nucleic acids or polypeptides also comprise at least one autocatalytic peptide cleavage site. In some alternatives, the at least one autocatalytic peptide cleavage site is a P2A autocatalytic peptide cleavage site. In some alternatives, the polypeptide encoded by the at least one CCHFV nucleic acid component or the at least one CCHFV polypeptide component are separated by the at least one autocatalytic peptide cleavage site.

[0012] In some alternatives, at least one M-segment strain sequence is provided in or encoded by the nucleic acids or polypeptides referenced above, such as a Gp38 (H) sequence, Gc ecto (H) sequence, or any combination thereof. In some alternatives, two M- segment sequences are provided in or encoded by the nucleic acids or polypeptides referenced thereof. In some alternatives, there is one M-segment sequence provided in or encoded by the nucleic acids or polypeptides. In some alternatives, the M-segment sequences are provided in or encoded by the nucleic acids or polypeptides in tandem. In some alternatives, the M-segment sequences are separated by peptide cleavage sites. In some alternatives, the M-segment sequences are separated from the S-segment sequences with an autocatalytic peptide cleavage site. In some alternatives, the autocatalytic peptide cleavage site is a P2A autocatalytic peptide cleavage site.

[0013] In some alternatives, the immunogenic compositions or product combinations comprise, consist essentially of, or consist of a nucleic acid, described above (e.g., any one or more of SEQ ID NO: IS, 21, 24, 27, 30, or 33), and a polypeptide, described above (e.g., any one or more of SEQ ID NO: 19, 22, 25, 28, 31, or 34 ). In some alternatives, the immunogenic compositions or product combinations are administered to a subject in a heterologous prime-boost approach. In some alternatives, the prime dose comprises the nucleic acid and the boost dose comprises the polypeptide. In some alternatives, the prime dose comprises any one or more of the aforementioned polypeptides and the boost dose comprises any one or more of the aforementioned nucleic acids. In some alternatives, the immunogenic compositions or product combinations are administered to a subject as a homologous prime- boost approach. In some alternatives, the prime dose comprises any one or more of the aforementioned nucleic acids and the boost dose comprises either the same nucleic acid or adifferent nucleic acid. In some alternatives, the prime dose comprises any one or more of the aforementioned polypeptides and the boost dose comprises either the same polypeptide or a different polypeptide. In some alternatives, the immunogenic compositions or product combinations further comprise an adjuvant. In some embodiments, the adjuvant is alum. In some alternatives, the nucleic acid is provided as a recombinant vector. In some alternatives, the recombinant vector is pVAXl. In some alternatives, the immunogenic compositions or product combinations are used for the prevention, treatment, amelioration, or inhibition of CCHFV or a sequela associated therewith in a subject, such as a mammal, preferably a human, which may, optionally, be selected or identified to receive a medicament for the prevention, treatment, amelioration, or inhibition of a CCHFV infection or a sequela associated therewith. Such subjects can be selected or identified by clinical evaluation or diagnostic evaluation or both.

[0014] Some alternatives described herein concern methods of generating an immune response in a subject, preferably a human, using the immunogenic compositions, product compositions, nucleic acids, or polypeptides described above (e.g., any one or more of SEQ ID NO: 18, 19, 21, 22, 24, 25, 27, 28, 30, 31, 33, or 34). In some alternatives, the methods comprise a heterologous prime-boost approach. In some alternatives, at least one prime dose is administered to the subject and at least one boost dose is administered to the subject. In some alternatives, the at least one prime dose is a nucleic acid. In some alternatives, the at least one boost dose is a polypeptide. In some alternatives, the at least one boost dose comprises an adjuvant, such as alum. In some alternatives, the at least one boost dose is administered at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, or 48 days or weeks after the at least one prime dose is administered or within a range of time defined by any two of the aforementioned time points. In some alternatives, the methods comprise a homologous prime-boost approach.

[0015] Additional alternatives concern an injection device comprising any one or more of the compositions described herein, such as any one or more of the nucleic acids or polypeptides set forth in any one or more of SEQ ID NO: 18, 19, 21, 22, 24, 25, 27, 28, 30, 31, 33, or 34. Such injection devices can comprise a single dose of such nucleic acid or polypeptide and such injection devices can have modified needle designs configured to enhance delivery of the nucleic acid or polypeptide or both. Such injection devices can be used with or without electroporation. Contemplated injections devices, which can include anyone or more of the nucleic acids or polypeptides of SEQ ID NO: 18, 19, 21, 22, 24, 25, 27, 28, 30, 31, 33, or 34 are described in U.S. Pat. App. Pub. No. 2016 / 0235928; PCT App. Pub. No. WO2014064534; U.S. Pat. Nos. 6,610,044; 6,132,419; 6,379,966; 6,897,068; 7,015,040; 7,214,369; 7,473,419; and 7,589,059, all of which are hereby expressly incorporated by reference in their entireties.

[0016] Preferred aspects of the present invention are related to the following numbered alternatives:

[0017] 1. A nucleic acid comprising at least one nucleic acid sequence encoding aCCHFV polypeptide and at least one nucleic acid sequence encoding a P2A autocatalytic polypeptide cleavage site.

[0018] 2. The nucleic acid of alternative 1, wherein the nucleic acid sequences encoding a CCHFV polypeptide comprise a nucleic acid sequence encoding an S-segment polypeptide NP (H), and a nucleic acid sequence encoding an M-segment polypeptide GP38 (H).

[0019] 3. The nucleic acid of any one of alternatives 1 or 2, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 21, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

[0020] 4. The nucleic acid of alternative 1, wherein the nucleic acid sequences encoding a CCHFV polypeptide comprise a nucleic acid sequence encoding an S-segment polypeptide NP (H), and a nucleic acid sequence encoding an M-segment polypeptide Gc_ecto (H).

[0021] 5. The nucleic acid of any one of alternatives 1 to 4, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 27, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

[0022] 6. The nucleic acid of alternative 2, further comprising a nucleic acid sequence encoding an Ig leader sequence.

[0023] 7. The nucleic acid of any one of alternatives 1 to 6, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homologyor sequence identity to SEQ ID NO: 18, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

[0024] 8. The nucleic acid of alternative 4, further comprising a nucleic acid sequence encoding an Ig leader sequence.

[0025] 9. The nucleic acid of any one of alternatives 1 to 8, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 24, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

[0026] 10. The nucleic acid of alternative 4, wherein the nucleic acid sequence encoding a CCHFV polypeptide further comprises a nucleic acid sequence encoding an M- segment polypeptide GP38 (H).

[0027] 11. The nucleic acid of any one of alternatives 1 to 10, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity7to SEQ ID NO: 33, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

[0028] 12. The nucleic acid of alternative 10, further comprising a nucleic acid sequence encoding an Ig leader sequence.

[0029] 13. The nucleic acid of any one of alternatives 1 to 12, wherein the nucleic acid has at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology pr sequence identity7to SEQ ID NO: 30, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

[0030] 14. A polypeptide comprising at least one CCITV polypeptide sequence and at least one P2A autocatalytic polypeptide cleavage site.

[0031] 15. The polypeptide of alternative 14, wherein the CCHFV polypeptide sequences comprise an S-segment NP (H) polypeptide sequence, and an M-segment GP38 (H) polypeptide sequence.

[0032] 16. The polypeptide of any one of alternatives 14 or 15, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 22, wherein the polypeptide comprises an epitope or antigen of CCHFV.

[0033] 17. The polypeptide of alternative 14, wherein the CCHFV polypeptide sequences comprise a S-segment NP (H) polypeptide sequence, and a Gc ecto (H) polypeptide sequence.

[0034] 18. The polypeptide of any one of alternatives 14 to 17, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 28, wherein the polypeptide comprises an epitope or antigen of CCHFV.

[0035] 19. The polypeptide of alternative 15, further comprising an Ig leader polypeptide sequence.

[0036] 20. The polypeptide of alternative 14 to 19, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 19, wherein the polypeptide comprises an epitope or antigen of CCHFV.

[0037] 21. The polypeptide of alternative 17, further comprising an Ig leader polypeptide sequence.

[0038] 22. The polypeptide of alternative 14 to 21, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 25, wherein the polypeptide comprises an epitope or antigen of CCHFV.

[0039] 23. The polypeptide of alternative 17, wherein the CCHFV polypeptide sequences further comprise an M-segment GP38 (H) polypeptide sequence.

[0040] 24. The polypeptide of alternative 14 to 23, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%, or 100% homology or sequence identity to SEQ ID NO: 34, wherein the polypeptide comprises an epitope or antigen of CCHFV.

[0041] 25. The polypeptide of alternative 23, further comprising an Ig leader polypeptide sequence.

[0042] 26. The polypeptide of alternative 14 to 25, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%, or 100% homology or sequence identity to SEQ ID NO: 31, wherein the polypeptide comprises an epitope or antigen of CCHFV.

[0043] 27. The nucleic acid of any one of alternatives 1 to 13 for use in a medicament, such as for the prevention, treatment or inhibition of CCHFV in a subject, preferably a human.

[0044] 28. The polypeptide of any one of alternatives 14 to 26 for use in a medicament, such as for the prevention, treatment or inhibition of CCHFV in a subject, preferably a human.

[0045] 29. The polypeptide of any one of alternatives 14 to 26 or 28, wherein the polypeptide is recombinantly expressed.

[0046] 30. The polypeptide of any one of alternatives 14 to 26 or 28 to 29, wherein the polypeptide is recombinantly expressed in a mammalian, bacterial, yeast, insect, or cell- free system.

[0047] 31. An immunogenic composition or product combination comprising:(a) a nucleic acid comprising at least one nucleic acid sequence encoding a CCHFV polypeptide; and(b) a polypeptide comprising at least one CCHFV polypeptide.

[0048] 32. The immunogenic composition or product combination of alternative31, wherein the at least one nucleic acid sequence encoding a CCHFV polypeptide comprises: i) a nucleic acid sequence encoding an S-segment NP (H) polypeptide; ii) a nucleic acid sequence encoding an M-segment GP38 (H) polypeptide; iii) a nucleic acid sequence encoding an M-segment Gc__ecto (H) polypeptide; iv) a nucleic acid sequence encoding an Ig leader polypeptide; v) a nucleic acid sequence encoding a P2A autocatalytic polypeptide cleavage site; or any combination thereof.

[0049] 33. The immunogenic composition or product combination of alternative 31 or 32, wherein the nucleic acid has at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 18, 21, 24, 27, 30, or 33, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV, which is optionally used in a medicament, such as for the prevention, treatment, or inhibition of CCHFV in a subject, such as a mammal, preferably a human.

[0050] 34. The immunogenic composition or product combination of any one of alternatives 31 to 33, wherein the nucleic acid is codon optimized for expression in a human.

[0051] 35. The immunogenic composition or product combination of any one of alternatives 31 to 34, wherein the at least one CCHFV polypeptide comprises: i) an S-segment NP (H) polypeptide sequence; ii) an M-segment GP38 (H) polypeptide sequence; lii) an M-segment Gc ecto (H) polypeptide sequence; iv) an Ig leader polypeptide sequence; v) a P2A autocatalytic polypeptide cleavage site sequence; or any combination thereof.

[0052] 36. The immunogenic composition or product combination of any one of alternatives 31 to 35, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 19, 22, 25, 28, 31, or 34, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV, which is optionally used in a medicament, such as for the prevention, treatment, or inhibition of CCHFV in a subject, such as a mammal, preferably a human.

[0053] 37. The immunogenic composition or product combination of any one of alternatives 31 to 36, wherein the polypeptide is recombinantly expressed.

[0054] 38. The immunogenic composition or product combination of any one of alternatives 31 to 37, wherein the polypeptide is recombinantly expressed in a mammalian, bacterial, yeast, insect, or cell-free system.

[0055] 39. The immunogenic composition or product combination of any one of alternatives 31 to 38, further comprising an adjuvant.

[0056] 40. The immunogenic composition or product combination of alternative39, wherein the adjuvant is alum.

[0057] 41. The immunogenic composition or product combination of any one of alternatives 31 to 40, wherein the nucleic acid is provided in a recombinant vector.

[0058] 42. A method of generating an immune response in a subject using the immunogenic composition or product combination set forth in any one of alternatives 31 to 41, comprising:a) administering to the subject at least one prime dose comprising the nucleic acid; and b) administering to the subject at least one boost dose comprising the polypeptide.

[0059] 43. The method of alternative 42, wherein the at least one boost dose further comprises an adjuvant.

[0060] 44. The method of alternative 43, wherein the adjuvant is alum.

[0061] 45. The method of any one of alternatives 42 to 44, wherein the at least one boost dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, or 48 days or weeks after the at least one prime dose is administered or within a range of time defined by any two of the aforementioned time points e.g., within 1-48 days or 1-48 weeks.

[0062] 46. The method of any one of alternatives 42 to 45, wherein the administration is provided enterally, orally, intranasally, parenterally, subcutaneously, intramuscularly, intradermally, or intravenously or any combination thereof, and optionally with in vivo electroporation.

[0063] 47. The method of any one of alternatives 42 to 46, wherein the administration is performed in conjunction with an antiviral therapy.

[0064] 48. The method of alternative 47, wherein the antiviral therapy comprises administration of chloroquine, hydroxychloroquine, favipiravir, favilavir, remdesivir, tocilizumab, galidesivir, sarilumab, lopinavir, ritonavir, darunavir, ribavirin, interferon- a, pegylated interferon-a, or interferon alfa-2b, or any combination thereof.

[0065] 49. An immunogenic composition or product combination for use in the treatment or inhibition of CCHFV, comprising:(a) a nucleic acid comprising at least one nucleic acid sequence encoding a CCHFV polypeptide; and(b) a polypeptide comprising at least one CCHFV polypeptide.

[0066] 50. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of alternative 49, wherein the at least one nucleic acid sequence encoding a CCHFV polypeptide comprises: i) a nucleic acid sequence encoding an S-segment NP (H) polypeptide ; ii) a nucleic acid sequence encoding an M-segment GP38 (H) polypeptide ;iii) a nucleic acid sequence encoding an M-segment Gc ecto (H) polypeptide ; iv) a nucleic acid sequence encoding an Ig leader polypeptide ; v) a nucleic acid sequence encoding a P2A autocatalytic polypeptide cleavage site; or any combination thereof.

[0067] 51. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of alternative 49 or 50, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 18, 21, 24, 27, 30, or 33, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV.

[0068] 52. The immunogenic composition or product combination for use in the treatment or inhibition of CCHF V of alternative 49 or 51, wherein the nucleic acid is codon optimized for expression in a human.

[0069] 53. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of alternatives 49 to 52, wherein the at least one CCHFV polypeptide comprises: i) an S-segment NP (H) polypeptide sequence; ii) an M-segment GP38 (H) polypeptide sequence; iii) an M-segment Gc ecto (H) polypeptide sequence; iv) an Ig leader polypeptide sequence; v) a P2A autocatalytic polypeptide cleavage site sequence; or any combination thereof.

[0070] 54. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of alternatives 49 to 53, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 19, 22, 25, 28, 31, or 34, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV.

[0071] 55. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of alternatives 49 to 54, wherein the polypeptide is recombinantly expressed.

[0072] 56. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of alternatives 49 to 55, wherein the polypeptide is recombinantly expressed in a mammalian, bacterial, yeast, insect, or cell-free system.

[0073] 57. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of alternatives 49 to 56, further comprising an adjuvant.

[0074] 58. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of alternative 57, wherein the adjuvant is alum.

[0075] 59. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of alternatives 49 to 58, wherein the nucleic acid is provided in a recombinant vector.BRIEF DESCRIPTION OF THE. DRAWINGS

[0076] In addition to the features described above, additional features and variations will be readily apparent from the following descriptions of the drawings and exemplary embodiments. It is to be understood that these drawings depict typical embodiments and are not intended to be limiting in scope.

[0077] FIGURE 1 depicts a schematic of the domains comprising the amino acid sequences for CCHFV immunogenic compositions SVF-003.1 -SVF-003.6.

[0078] FIGURE 2 depicts nucleotide sequences for BamHI, Xbal, Kozak sequence, and stop codon, and ammo acid and nucleotide sequences for the IgE leader, P2A autocleavage site, GP38_Hoti, Gc_ecto Hoti, NP_Hoti, and additional cleavage site.

[0079] FIGURE 3A-B depict schematics of the glycoprotein precursors in wild type IbArl0200 (FIG. 3 A) and Hoiti (FIG. 3B) strains.

[0080] FIGURE 4 depicts the schematic, nucleotide, and amino acid information for SAT-003.1 CCHFV immunogenic compositions.

[0081] FIGURE 5 depicts the schematic, nucleotide, and ammo acid information for SVF-003.2 CCHFV immunogenic compositions.

[0082] FIGURE 6 depicts the schematic, nucleotide, and ammo acid information for SVF-003.3 CCHFV immunogenic compositions.

[0083] FIGURE 7 depicts the schematic, nucleotide, and ammo acid information for SVF-003.4 CCHFV immunogenic compositions.

[0084] FIGURE 8 depicts the schematic, nucleotide, and ammo acid information for SVF-OO3.5 CCHFV immunogenic compositions.

[0085] FIGURE 9 depicts the schematic, nucleotide, and ammo acid information for SVF-003.6 CCHFV immunogenic compositions.

[0086] FIGURE 10A-10D depict a study to assess the immunogenic effects of the CCHFV immunogenic compositions SVF-003.1 -SVF-003.6. 10A depicts results for plasmid preparation of the DNA immunogenic compositions. 10B depicts dose amounts and injection volumes for the immunogenic compositions and controls used in the prime / boost strategy of the study. 10C and 10D depict anti-N antibody titers in immunized mice at two weeks after immunization with the immunogenic DNA compositions.

[0087] FIGURE 11 A-l IB depict anti-N antibody titers in immunized mice at week five, initial immunization with the immunogenic DNA compositions (prime) was performed at week zero, and immunization with the immunogenic protein compositions (boost) was performed at week three.

[0088] FIGURE 12A-12H depict the results of an assessment of IFNy production by T cells in response to CCHFV antigens by ELISpot Proportions of IFN-y+ and IL-2+ T cells responding to the peptides of the CCHFV peptide pool were measured for mice immunized with SVF.003.1 -SVF.003.6 and controls.DETAILED DESCRIPTION

[0089] In the following detailed description, reference is made to the accompanying drawings, which form a part hereof. In the drawings, similar symbols typically identify similar components, unless context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described herein, and illustrated in the Figures,can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are explicitly contemplated herein.Definitions

[0090] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary’ skill in the art. All patents, applications, published applications and other publications referenced herein are expressly incorporated by reference in their entireties unless stated otherwise. In the event that there are a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

[0091] The articles “a” and “an” are used herein to refer to one or to more than one (for example, at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0092] The terms “about” or “around” as used herein refer to a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length that varies by as much as 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1% to a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length.

[0093] Throughout this specification, unless the context requires otherwise, the words “comprise,” “comprises,” and “comprising” will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.

[0094] By “consisting of” is meant including, and limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of’ indicates that the listed elements are required or mandatory, and that no other elements may be present. By “consisting essentially of” is meant including any elements listed after the phrase and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase “consisting essentially of” indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present depending upon whether or not they materially affect the activity or action of the listed elements.

[0095] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which thisdisclosure belongs. If there is a plurality of definitions for a term herein, those in this section prevail unless stated otherwise. The practice of the present disclosure wall employ, unless indicated specifically to the contrary, conventional methods of molecular biology and recombinant DNA techniques within the skill of the art, many of which are described below7for the purpose of illustration.

[0096] The terms “individual”, “subject”, or “patient” as used herein, means a human or a non-human mammal, e.g., a dog, a cat, a mouse, a rat, a cow7, a sheep, a pig, a goat, a non-human primate, or a bird, e.g., a chicken, as w7ell as any other vertebrate or invertebrate.

[0097] The term “mammal” is used in its usual biological sense. Thus, it specifically includes, but is not limited to, primates, including simians (chimpanzees, apes, monkeys) and humans, cattle, horses, sheep, goats, sw'ine, rabbits, dogs, cats, rodents, rats, mice, guinea pigs, or the like.

[0098] Some embodiments described herein relate to pharmaceutical compositions that comprise, consist essentially of, or consist of an effective amount of an oligonucleotide described herein and a pharmaceutically acceptable carrier, excipient, or combination thereof, A pharmaceutical composition described herein is suitable for human and / or veterinary applications,

[0099] The terms “function” and “functional” as used herein refer to a biological, enzymatic, or therapeutic function,

[0100] The terms “effective amount” or “effective dose” is used to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal response indicated. For example, an effective amount, of compound can be the amount needed to alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated This response may occur in a tissue, system, animal or human and includes alleviation of the signs or symptoms of the disease being treated. Determination of an effective amount is well within the capability of those skilled in the art, in view of the disclosure provided herein. The effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated, and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize.

[0101] The term “pharmaceutically acceptable salts” includes relatively non-toxic, inorganic and organic acid, or base addition salts of compositions, including without limitation, analgesic agents, therapeutic agents, other materials, and the like. Examples of pharmaceutically acceptable salts include those derived from mineral acids, such as hydrochloric acid and sulfuric acid, and those derived from organic acids, such as ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. Examples of suitable inorganic bases for the formation of salts include phosphates, hydroxides, carbonates, and bicarbonates of ammonia, sodium, lithium, potassium, calcium, magnesium, aluminum, zinc, and the like. Salts may also be formed with suitable organic bases, including those that are non-toxic and strong enough to form such salts. For example, the class of such organic bases may include but are not limited to mono-, di-, and trialkylamines, including methylamine, dimethylamine, and triethylamine; mono-, di-, or trihydroxyalkylamines including mono-, di-, and triethanolamine; amino acids, including glycine, arginine and lysine; guanidine; N-methylglucosamine; N-methylglucamine; L-glutamine; N-methylpiperazine; morpholine; ethylenediamine; or N-benzylphenethylamine; tri hydroxymethyl aminoethane.

[0102] “Formulation”, “pharmaceutical composition”, and “composition” as used interchangeably herein are equivalent terms referring to a composition of matter for administration to a subject.

[0103] The term “pharmaceutically acceptable” means compatible with therapy for a subject, and in particular, a human.

[0104] The terms “agent” refers to an active agent that has biological activity and may be used in a therapy. Also, an “agent” can be synonymous with “at least one agent,” “compound,” or “at least one compound,” and can refer to any form of the agent, such as a derivative, analog, salt or a prodrug thereof. The agent can be present in various forms, components of molecular complexes, and pharmaceutically acceptable salts (e.g., hydrochlorides, hydrobromides, sulfates, phosphates, nitrates, borates, acetates, maleates, tartrates, or salicylates). The term “agent” can also refer to any pharmaceutical molecules or compounds, therapeutic molecules or compounds, matrix forming molecules or compounds, polymers, synthetic molecules and compounds, natural molecules and compounds, and any combination thereof.

[0105] Proper formulation is dependent upon the route of administration chosen. Techniques for formulation and administration of the compounds described herein are known to those skilled in the art. Multiple techniques of administering one or more of the nucleic acids or polypeptides described herein or immunogenic compositions comprising said nucleic acids or polypeptides described herein are contemplated including, but not limited to, enteral, oral, rectal, topical, sublingual, buccal, intraaural, epidural, epicutaneous, aerosol, parenteral, intramuscular, subcutaneous, intra-arterial, intravenous, intraportal, intra-articular, intradermal, peritoneal, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intranasal or intraocular injections. Pharmaceutical compositions will generally be tailored to the specific intended route of administration. The pharmaceutical compositions described herein can also be administered to subjects along with other therapies, such as T cells, Natural Killer cells, B cells, macrophages, lymphocytes, stem cells, bone marrow cells, or hematopoietic stem cells.

[0106] The term “isolated” as used herein refers to material that is substantially or essentially free from components that normally accompany it in its native state. For example, an “isolated cell,” as used herein, includes a cell that has been purified from the milieu or organisms in its naturally occurring state, a cell that has been removed from a subject or from a culture, for example, it is not significantly associated with in vivo or in vitro substances.

[0107] The pharmaceutical compound can also be administered in a local rather than systemic manner, for example, via injection of the compound directly into an organ, tissue, or infected area, often in a depot or sustained release formulation.

[0108] The pharmaceutical compositions disclosed herein may be manufactured in a manner that is itself known, e.g., by mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tableting processes. As described herein, compounds used in a pharmaceutical composition may be provided as salts with pharmaceutically compatible counterions.

[0109] As used herein, a “carrier” refers to a compound, particle, solid, semi-solid, liquid, or diluent that facilitates the passage, delivery and / or incorporation of a compound to cells, tissues and / or bodily organs. For example, without limitation, a lipid nanoparticle (LNP) is a type of carrier that can encapsulate an oligonucleotide to thereby protect theoligonucleotide from degradation during passage through the bloodstream and / or to facilitate deliver}' to a desired organ, such as to the liver.

[0110] As used herein, a “diluent” refers to an ingredient in a pharmaceutical composition that lacks pharmacological activity but may be pharmaceutically necessary or desirable. For example, a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and / or administration. It may also be a liquid for the dissolution of a drug to be administered by injection, ingestion or inhalation. A common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the composition of human blood.

[0111] The term “excipient” has its ordinary meaning as understood in light of the specification, and refers to inert substances, compounds, or materials added to a pharmaceutical composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability etc., to the composition. Excipients with desirable properties include but are not limited to preservatives, adjuvants, stabilizers, solvents, buffers, diluents, solubilizing agents, detergents, surfactants, chelating agents, antioxidants, alcohols, ketones, aldehydes, ethylenediaminetetraacetic acid (EDTA), citric acid, salts, sodium chloride, sodium bicarbonate, sodium phosphate, sodium borate, sodium citrate, potassium chloride, potassium phosphate, magnesium sulfate sugars, dextrose, dextran, fructose, mannose, lactose, galactose, sucrose, sorbitol, cellulose, methyl cellulose, hydroxypropyl methyl cellulose (hypromellose), glycerin, polyvinyl alcohol, povidone, propylene glycol, serum, amino acids, polyethylene glycol, polysorbate 20, polysorbate 80, sodium deoxy cholate, sodium taurodeoxy cholate, magnesium stearate, octylphenol ethoxylate, benzethonium chloride, thimerosal, gelatin, esters, ethers, 2-phenoxy ethanol, urea, or vitamins, or any combination thereof. The amount of the excipient may be found in a pharmaceutical composition at a percentage of 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 100% w / w or any percentage by weight in a range defined by any two of the aforementioned numbers.

[0112] The term “adjuvant” as used herein refers to a substance, compound, or material that stimulates the immune response and increase the efficacy of protective immunity and is administered in conjunction with an immunogenic antigen, epitope, or composition.Adjuvants serve to improve immune responses by enabiing a continual release of antigen, up- regulation of cytokines and chemokines, cellular recruitment at the site of administration, increased antigen uptake and presentation in antigen presenting cells, or activation of antigen presenting cells and inflammasomes. Commonly used adjuvants, which can be included in any one or more of the formulations set forth herein include but are not limited to alum, aluminum salts, aluminum sulfate, aluminum hydroxide, aluminum phosphate, calcium phosphate hydroxide, potassium aluminum sulfate, oils, mineral oil, paraffin oil, oil-in-water emulsions, detergents, MF59©, squalene, AS03, a-tocopherol, polysorbate 80, AS04, monophosphoryl lipid A, virosomes, nucleic acids, polyinosinic:polycytidylic acid, saponins, QS-21, proteins, flagellin, cytokines, chemokines, IL-1, IL-2, IL- 12, IL- 15, IL-21, imidazoquinolines, CpG oligonucleotides, lipids, phospholipids, dioleoyl phosphatidylcholine (DOPC), trehalose dimycolate, peptidoglycans, bacterial extracts, lipopolysaccharides, or Freund’s Adjuvant, or any combination thereof.

[0113] The term “purity” of any given substance, compound, or material as used herein refers to the actual abundance of the substance, compound, or material relative to the expected abundance. For example, the substance, compound, or material may be at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% pure, including all decimals in between. Purity may be affected by unwanted impurities, including but not limited to side products, isomers, enantiomers, degradation products, solvent, carrier, vehicle, or contaminants, or any combination thereof. Purity can be measured technologies including but not limited to chromatography, liquid chromatography, gas chromatography, spectroscopy, UV-visible spectrometry, infrared spectrometry, mass spectrometry, nuclear magnetic resonance, gravimetry, or titration, or any combination thereof.

[0114] Some embodiments disclosed herein related to selecting a subject or patient in need of receiving one or more of the nucleic acids or polypeptides described herein. In some embodiments, a patient or subject is selected who is in need of treatment or inhibition of a viral infection caused by Crimean Congo hemorrhagic fever virus (CCHFV) or a sequela associated therewith. In some embodiments, a patient or subject is selected who has previously received a therapy for CCHFV. In some embodiments, a patient or subject is selected who has previously received a therapy for being at risk of CCHFV infection or a sequela associated therewith. In some embodiments, a patient or subject is selected who has developed arecurrence of CCHFV infection. In some embodiments, a patient or subject is selected who has developed resistance to therapies for CCHFV infection or a sequela associated therewith. In some embodiments, a patient or subject is selected who may have any combination of the aforementioned selection criteria. Such selections can be made by clinical and diagnostic evaluation of the subject or a combination of both.

[0115] The terms “treat”, “treating”, “treatment”, “therapeutic”, or “therapy” as used herein has its ordinary meaning as understood in light of the specification, and do not necessarily mean total cure or abolition of the disease or condition. The term “treating” or “treatment” as used herein (and as well understood in the art) also means an approach for obtaining beneficial or desired results in a subject’s condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of the extent of a disease, stabilizing (i.e., not worsening) the state of disease, prevention of a disease's transmission or spread, delaying or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable. “Treating” and “treatment” as used herein also include prophylactic treatment. Treatment methods comprise administering to a subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may comprise a series of administrations. The compositions are administered to the subject in an amount and for a duration sufficient to treat the patient. The length of the treatment period depends on a variety of factors, such as the severity of the condition, the age and genetic profile of the patient, the concentration of active agent, the activity of the compositions used in the treatment, or a combination thereof. It will also be appreciated that the effective dosage of an agent used for the treatment or prophylaxis may increase or decrease over the course of a particular treatment or prophylaxis regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. The term “prophylactic treatment” refers to treating a subject who does not yet exhibit symptoms of a disease or condition, but who is susceptible to, or otherwise at risk of, a particular disease or condition, whereby the treatment reduces the likelihood that the patient wall develop the disease or condition. Theterm “therapeutic treatment” refers to administering treatment to a subject already suffering from or developing a disease or condition.

[0116] The term “inhibit” as used herein has its ordinary meaning as understood in light of the specification, and may refer to the reduction or prevention of a viral infection, such as CCHFV. The reduction can be by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or an amount that is within a range defined by any two of the aforementioned values. As used herein, the term “delay” has its ordinary meaning as understood in light of the specification, and refers to a slowing, postponement, or deferment of an event, such as a viral infection, to a time which is later than would otherwise be expected. The delay can be a delay of 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or an amount within a range defined by any two of the aforementioned values. The terms inhibit and delay may not necessarily indicate a 100% inhibition or delay. A partial inhibition or delay may be realized.

[0117] The term “immunogenic composition” as used herein refers to a substance or mixture of substances, including but not limited to antigens, epitopes, nucleic acids, peptides, polypeptides, proteins, polysaccharides, lipids, haptens, toxoids, inactivated organisms, or attenuated organisms, or any combination thereof, intended to elicit an immune response when administered to a host. The immune response includes both an innate and adaptive immune response, the latter of which establishes a lasting immunological memory through cells such as memory T cells and memory B cells. The antibodies created during the initial immune response to the immunogenic composition can be produced in subsequent challenges of the same antigens, epitopes, nucleic acids, peptides, polypeptides, proteins, polysaccharides, lipids, haptens, toxoids, inactivated organisms, or atenuated organisms, or a live organism or pathogen that exhibits the antigens, epitopes, nucleic acids, peptides, polypeptides, proteins, polysaccharides, lipids, haptens, or toxoids or any combination thereof. In this manner, the immunogenic composition may serve as a vaccine against a specific pathogen. Immunogenic compositions may also include one or more adjuvants to stimulate the immune response and increase the efficacy of protective immunity.

[0118] The term “product combination” as used herein refers to set of two or more individual compounds, substances, materials, or compositions that can be used together for a unified function. In some embodiments, a product combination comprises at least one nucleic acid composition and at least one polypeptide composition that are used together to elicit animmune response when administered to a host, optionally to a greater degree than would be elicited if only one composition type were to be administered.

[0119] The terms “nucleic acid” or “nucleic acid molecule” as used herein refers to polynucleotides, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), oligonucleotides, fragments generated by the polymerase chain reaction (PCR), and fragments generated by any of ligation, scission, endonuclease action, and exonuclease action. Nucleic acid molecules can be composed of monomers that are naturally-occurring nucleotides (such as DNA and RNA), or analogs of naturally-occurring nucleotides (e.g., enantiomeric forms of naturally-occurring nucleotides), or a combination of both. Modified nucleotides can have alterations in sugar moieties and / or in pyrimidine or purine base moieties. Sugar modifications include, for example, replacement of one or more hydroxyl groups with halogens, alkyl groups, amines, and azido groups, or sugars can be functionalized as ethers or esters. Moreover, the entire sugar moiety can be replaced with sterically and electronically similar structures, such as aza-sugars and carbocyclic sugar analogs. Examples of modifications in a base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well- known heterocyclic substitutes. Nucleic acid monomers can be linked by phosphodi ester bonds or analogs of such linkages. Analogs of phosphodiester linkages include phosphor othioate, phosphorodithi oate, phosphoroselenoate, phosphorodisel enoate, phosphoroanil othioate, phosphoranilidate, or phosphoramidate. The term “nucleic acid molecule” also includes so- called “peptide nucleic acids,” which comprise naturally-occurring or modified nucleic acid bases attached to a polyamide backbone. Nucleic acids can be either single stranded or double stranded. “Oligonucleotide” can be used interchangeable with nucleic acid and can refer to either double stranded or single stranded DNA or RNA. A nucleic acid or nucleic acids can be contained in a nucleic acid vector or nucleic acid construct (e.g. plasmid, virus, bacteriophage, cosmid, fosmid, phagemid, bacterial artificial chromosome (BAC), yeast, artificial chromosome (YAC), or human artificial chromosome (HAC)) that can be used for amplification and / or expression of the nucleic acid or nucleic acids in various biological systems. Typically, the vector or construct will also contain elements including but not limited to promoters, enhancers, terminators, inducers, ribosome binding sites, translation initiation sites, start codons, stop codons, polyadenylation signals, origins of replication, cloning sites, multiple cloning sites, restriction enzyme sites, epitopes, reporter genes, selection markers,antibiotic selection markers, targeting sequences, peptide purification tags, or accessory genes, or anv combination thereof.

[0120] A nucleic acid or nucleic acid molecule can comprise one or more sequences encoding different peptides, polypeptides, or proteins. These one or more sequences can be joined in the same nucleic acid or nucleic acid molecule adjacently, or with extra nucleic acids in between, e.g. linkers, repeats or restriction enzyme sites, or any other sequence that is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 bases long, or any length in a range defined by any two of the aforementioned lengths. The term “downstream” on a nucleic acid as used herein refers to a sequence being after the 3 ’-end of a previous sequence, on the strand containing the encoding sequence (sense strand) if the nucleic acid is double stranded. The term “upstream” on a nucleic acid as used herein refers to a sequence being before the 5 ’-end of a subsequent sequence, on the strand containing the encoding sequence (sense strand) if the nucleic acid is double stranded. The term “grouped” on a nucleic acid as used herein refers to two or more sequences that occur in proximity either directly or with extra nucleic acids in between, e.g. linkers, repeats, or restriction enzyme sites, or any other sequence that is 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 bases long, or any length in a range defined by any two of the aforementioned lengths, but generally not with a sequence in between that encodes for a functioning or catalytic polypeptide, protein, or protein domain.

[0121] The term “codon optimized” regarding a nucleic acid as used herein refers to the substitution of codons of the nucleic acid to enhance or maximize translation in a host of a particular species without changing the polypeptide sequence based on species -specific codon usage biases and relative availability of each aminoacyl-tRNA in the target cell cytoplasm. Codon optimization and techniques to perform such optimization is known in the art. Programs containing algorithms for codon optimization are known to those skilled in the art. Programs can include, for example, OptimumGene, GeneGPS® algorithms, etc. Additionally, synthetic codon optimized sequences can be obtained commercially for example from Integrated DNA Technologies and other commercially available DNA sequencing services. Those skilled in the art will appreciate that gene expression levels are dependent on many factors, such as promoter sequences and regulatory elements. As noted for most bacteria,small subsets of codons are recognized by tRNA species leading to translational selection, which can be an important limit on protein expression. In this aspect, many synthetic genes can be designed to increase their protein expression level.

[0122] The nucleic acids described herein comprise nucleobases. Primary, canonical, natural, or unmodified bases are adenine, cytosine, guanine, thymine, and uracil. Other nucleobases include but are not limited to purines, pyrimidines, modified nucleobases, 5-methylcytosine, pseudouridine, dihydrouridine, inosine, 7 -methylguanosine, hypoxanthine, xanthine, 5,6-dihydrouracil, 5-hydroxymethylcytosine, 5-bromouracil, isoguanine, isocytosine, aminoallyl bases, dye-labeled bases, fluorescent bases, or biotin-labeled bases.

[0123] The terms “peptide”, “polypeptide”, and “protein” as used herein refers to macromolecules comprised of amino acids linked by peptide bonds. The numerous functions of peptides, polypeptides, and proteins are known in the art, and include but are not limited to enzymes, structure, transport, defense, hormones, or signaling. Peptides, polypeptides, and proteins are often, but not always, produced biologically by a ribosomal complex using a nucleic acid template, although chemical syntheses are also available. By manipulating the nucleic acid template, peptide, polypeptide, and protein mutations such as substitutions, deletions, truncations, additions, duplications, or fusions of more than one peptide, polypeptide, or protein can be performed. These fusions of more than one peptide, polypeptide, or protein can be joined in the same molecule adjacently, or with extra amino acids in between, e.g. linkers, repeats, epitopes, or tags, or any other sequence that is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 300 bases long, or any length in a range defined by any two of the aforementioned lengths. The term “downstream” on a polypeptide as used herein refers to a sequence being after the C-terminus of a previous sequence. The term “upstream” on a polypeptide as used herein refers to a sequence being before the N-terminus of a subsequent sequence.

[0124] In some embodiments, the nucleic acid or peptide sequences presented herein and used in the examples are functional in various biological systems including but not limited to humans, mice, rabbits, E. coll, yeast, and mammalian cells. In other embodiments, nucleic acid or peptide sequences sharing 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% similarity, sequenceidentity, or homology, or any percentage within a range defined by any two of the aforementioned percentages similarity to the nucleic acid or peptide sequences presented herein and used in the examples can also be used with no effect on the function of the sequences in biological systems. As used herein, the term “similarity” refers to a nucleic acid or peptide sequence having the same overall order of nucleotide or ammo acids, respectively, as a template nucleic acid or peptide sequence with specific changes such as substitutions, deletions, repetitions, or insertions within the sequence. In some embodiments, two nucleic acid sequences sharing as low as 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% similarity can encode for the same polypeptide by comprising different codons that encode for the same ammo acid during translation.

[0125] The term “recombinantly expressed” as used herein refers to the production of proteins in optimized or adapted biological systems. These systems provide advantages over protein expression in a natural host, including but not limited to high expression (overexpression), ease of purification, ease of transformation, inducibility, low cost, or stability of the protein. In some embodiments, proteins are expressed in mammalian, bacteria, yeast, insect, or cell-free recombinant expression systems. Each system has its own advantages or disadvantages. For example, bacterial expression systems are highly optimized for overexpression, but may cause misfolding or aggregation of the produced protein, yeast systems are useful when post-translational modifications are necessary, and insect and mammalian systems are useful for proper RNA splicing that occurs in higher-order organisms. In some embodiments, recombinant polypeptides are produced and purified from mammalian, human, primary, immortalized, cancer, stem, fibroblasts, human embryonic kidney (HEK) 293, Chinese Hamster Ovary (CHO), bacterial, Escherichia coli, yeast, Saccharomyces cerevisiae, Pichia pasioris, insect, Spodoptera frugiperda Sf9, or S. frugiperda 5 / 21 cells, or in a cell-free system. In some embodiments, expression genes, vectors, or constructs are delivered to the recombinant expression systems m the form of plasmids, bacteriophages, viruses, adeno-associated viruses (AAVs), baculovirus, cosmids, fosmids, phagemids, BACs, YACs, or HACs. For more discussion on recombinant expression systems, see Gomes et al. “An Overview of Heterologous Expression Host Systems for the Production of Recombinant Proteins” ((2016) Adv. Anim. Vet. Sci. 4(7):346-356), hereby expressly incorporated by reference in its entirety.

[0126] The terms “autocatalytic peptide cleavage site” or “2A peptide” as used herein refer to a peptide sequence that undergo cleavage of a peptide bond between two constituent ammo acids, resulting in separation of the two proteins that flank the sequence. The cleavage is believed to be a result of a ribosomal “skipping” of the peptide bond formation between the C-terminal proline and glycine in the 2A peptide sequence. Four autocatalytic peptide cleavage site sequences identified to date have seen substantial use in biomedical research: foot-and-mouth disease virus 2A (F2A); equine rhinitis A virus (ERAV) 2A (E2A); porcine teschovirus- 1 2A (P2A), and Thosea asigna virus 2A (T2A). In some embodiments, the P2A autocatalytic peptide cleavage site nucleic acid (SEQ ID NO: 9) and polypeptide (SEQ ID NO: 8) sequences are used. In some embodiments, the P2A nucleic acid or polypeptide used can be substituted with an F2A, E2A, or T2A nucleic acid or polypeptide.

[0127] In some embodiments, the nucleic acids or peptides used herein comprise sequences that produce CCHFV antigens. CCHFV is an enveloped negative stranded RNA virus with a helical capsid symmetry' belonging to the Bunyaviruses that causes a severe and often lethal hemorrhagic disease. It has been shown that the M and N gene segments are strongly associated with immune responses that protect against disease. In addition, immune responses against the GP38 protein, encoded by the M segment, has shown protective properties in the absence of neutralizing antibodies. In some embodiments, highly potent chimeric genes containing sequence from both envelope and nucleoproteins are provided that induce both neutralizing antibodies and T cells and effectively protect mice against infection and / or disease in various models. In some embodiments, the M-segment sequence used is GP38 (H). In some embodiments, the GP38 (H) nucleic acid (SEQ ID NO: 11) sequence and polypeptide sequence (SEQ ID NO: 10) are used. In some embodiments, the M-segment sequence used is Gc_ecto (H). In some embodiments, the Gc__ecto (H) nucleic acid (SEQ ID NO: 13) sequence and polypeptide sequence (SEQ ID NO: 12) are used. In some embodiments, the S-segment sequence used is NP (H). In some embodiments, the NP (H) nucleic acid (SEQ ID NO: 15) sequence and polypeptide sequence (SEQ ID NO: 14) are used.

[0128] In some embodiments, the sequence further includes an IgE leader sequence. Tthe term “IgE leader sequence” as used herein refers to the amino acid sequence MDWTWILFLVAAATRVHS (SEQ ID NO: 6), which can be appended to the N-terminus ofa protein to both enhance translation and increase immunogenicity. Translation is particularly upregulated when the IgE leader sequence is used in combination with a functional Kozak sequence. An exemplary embodiment of a nucleic acid sequence that encodes for the ammo acid IgE leader sequence is represented as SEQ ID NO: 7. However, it would be clearly apparent to one skilled in the art to develop alternative nucleic acid sequence that would result in the same amino acid sequence when translated. Additional insight into the use of an IgE leader sequence may be found in Vijayachari et al. “Immunogenicity of a novel enhanced consensus DNA vaccine encoding the leptospiral protein LipL45” Hum. Vaccm. Immunother. (2015); 11 (8): 1945-53, which is hereby expressly incorporated by reference in its entirety.Methods of Therapy

[0129] The terms “prime” and “boost” as used herein related to separate immunogenic compositions used in a heterologous prime-boost immunization approach. Immunizations or vaccines commonly require more than one administration of an immunogenic composition to induce a successful immunity against a target pathogen in a host. Compared to this homologous approach where the same composition is provided for all administrations, a heterologous prime-boost administration can be more effective in establishing robust immunity with greater antibody levels and improved clearing or resistance against some pathogens such as CCHFV. In a heterologous prime-boost administration, at least one prime dose comprising one type of immunogenic composition is first provided. After the at least one prime dose is provided, at least one boost dose comprising another type of immunogenic composition is then provided. Administration of the at least one boost dose is performed at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, or 48 days or weeks after the at least one prime dose is administered or within a range of time defined by any two of the aforementioned time points e.g., within 1 -48 days or 1-48 weeks. In some embodiments, the prime dose comprises a nucleic acid (e.g. DNA or RNA) that encodes for one or more antigens or epitopes, and the boost dose comprises a polypeptide that comprises one or more antigens or epitopes. In the host, the nucleic acid prime is translated in vivo to elicit an immune reaction and causes a greater response against the subsequent polypeptide boost. In some embodiments, the nucleic acid prime comprises, consists essentially of, or consists of sequences from CCHFV In some embodiments, the sequences from CCHFV encode for an S-segment and / or M-segment polypeptide. In some embodiments, the nucleic acid prime also includes at least one Ig leader sequence. In some embodiments, the nucleic acid sequences are codon optimized for expression in humans. In some embodiments, the polypeptide boost comprises, consists essentially of, or consists of polypeptides from CCHFV. In some embodiments, the polypeptides from CCHFV are S-segment and / or M-segment polypeptides. In some embodiments, the prime dose is a polypeptide, and the boost dose is a nucleic acid. General information about heterologous prime-boost approaches can be found in PCT Publications WO 2006 / 013106, WO 2006 / 040334, WO 2008 / 094188, each of which are hereby expressly incorporated by reference for the purpose of describing prime-boost methods.

[0130] In some embodiments, administration of the nucleic acid prime and polypeptide boost comprising components of CCHFV in a subject (e.g. mouse, rabbit, monkey, human) results in greater anti-S-segment, anti-M-segment, anti- CCHFV antibody titer at a ratio of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 50, 100, 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 5000, 10000, 100000, or 1000000 or any ratio within a range defined by any two of the aforementioned ratios compared to nucleic acid-only or polypeptide-only immunized, or unimmunized control organisms, quantified by techniques known in the art such as ELISA, In some embodiments, administration of the nucleic acid prime and polypeptide boost comprising components of CCHFV in a subject results in serum that neutralizes the in vitro or in vivo infectivity of CCHFV more effectively and reduces the incidence of infection or multiplicity of infection (MOI) to a ratio of 0.00001, 0.00005, 0.0001 , 0.0005, 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0,7, 0.8, 0.9, or 1.0 or any ratio within a range defined by any two of the aforementioned ratios compared to sera from nucleic acid-only or polypeptide-only immunized, or unimmunized control organisms. In some embodiments, administration of the nucleic acid prime and polypeptide boost comprising components of CCHFV in a subject results in a greater number of interferon gamma (IFNy)- positive cells (e.g. T cells, macrophages, natural killer (NK) cells) at a ratio of 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 5000, or 10000, or any ratio within a range defined by any two of the aforementioned ratios compared to nucleic acid-only or polypeptide-only immunized, or unimmunized control organisms.

[0131] In some embodiments, the immunogenic compositions or product combinations are administered with an adjuvant. In some embodiments, the immunogenic compositions or product combinations are administered enterally, orally, intranasally, parenterally, subcutaneously, intramuscularly, intradermally, or intravenously or any combination thereof. In some embodiments, the immunogenic compositions or product combinations are administered in conjunction with an antiviral therapy compound known or hypothesized have an effect against CCHFV, or side effects caused by CCHFV, including but not limited to chloroquine, hydroxychloroquine, favipiravir, favilavir, remdesivir, tocilizumab, galidesivir, sarilumab, lopmavir, ritonavir, darunavir, ribavirin, interferon-a, pegylated interferon-a, or interferon alfa-2b, or any combination thereof.

[0132] The terms “m vivo electroporation”, “electroporation”, and “EP” as used herein refers to the deliver}' of genes, nucleic acids, DNA, RNA, proteins, or vectors into cells of living tissues or organisms using electrical currents using techniques known in the art. Electroporation can be used as an alternative to other methods of gene transfer such as viruses (transduction), lipofection, gene gun (biolistics), microinjection, vesicle fusion, or chemical transformation. Electroporation limits the risk of immunogenicity and detrimental integration or mutagenesis of the cell genome. DNA vectors such as plasmids are able to access the cell nucleus, enabling transcription and translation of constituent genes. In some embodiments, the genes, nucleic acids, DNA, RNA, proteins, or vectors are added to the target tissue or organism by subcutaneous, intramuscular, or intradermal injection. An electroporator then delivers short electrical pulses via electrodes placed within or proximal to the injected sample. As used herein, the term “im / EP” refers to in vivo electroporation of a sample delivered intramuscularly (“im”).

[0133] The term “transient immunosuppression” as used herein refers to a C57BL / 6 mouse model wherein transient immunosuppression is accomplished by administration of type I interferon (IFN-1) blocking antibody (mAb)-5A3 before challenge. The term “type I Interferon receptor (IFNAR‘ / _) mice” refers to a mouse model wherein the mice are engineered not to express the type I Interferon receptor. Other similar mouse models can be used as alternatives.

[0134] Where a range of values is provided, it is understood that the upper and lower limit, and each intervening value between the upper and lower limit of the range is encompassed within the embodiments.

[0135] The term “% w / w” or “% wt / wt” as used herein has its ordinary meaning as understood in light of the specification and refers to a percentage expressed in terms of the weight of the ingredient or agent over the total weight of the composition multiplied by 100. The term “% v / v” or “% voi / vol” as used herein has its ordinary meaning as understood in the light of the specification and refers to a percentage expressed in terms of the liquid volume of the compound, substance, ingredient, or agent over the total liquid volume of the composition multiplied by 100.

[0136] The invention is generally disclosed herein using affirmative language to describe the numerous embodiments. The invention also includes embodiments in which subject matter is excluded, in full or in part, such as substances or materials, method steps and conditions, protocols, or procedures.EXAMPLES

[0137] Some aspects of the embodiments discussed above are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the present disclosure. Those in the art will appreciate that many other embodiments also fall within the scope of the invention, as it is described herein above and in the claims.Example 1 : Exemplary CCHFV immunogenic constructs

[0138] The recombinant CCHFV constructs were assembled by combining S- segment sequence NP (H), and one or more M-segment sequences (GP38 (H) and Gc_ecto (IT)) with a P2A autocatalytic peptide cleavage site, and / or an Ig leader sequence, and / or an RSKR cleavage site. Schematics for six exemplary recombinant constructs are shown in FIG. 1, and nucleotide and ammo acid sequences comprising the constructs are depicted in FIG. 2, and FIGs. 4-9. Alternatively, nucleotide and amino acid sequences for CCHFV constructs are also included in TABLE 1. Additionally, the glycoprotein precursors for IbAr 10200 and Hoiti and associated cleavage sites are illustrated in FIG. 3A-B.TABLE 1Example 2: Evaluation of Immunogenic Effect of CCHFV Vaccine Compositions

[0139] The immunogenic effects of the CCHFV immunogenic compositions were tested by immunizing mice with the CCHFV immunogenic compositions comprising the protein and nucleotide sequences of SVF-003.1, SVF-003.2, SVF-003.3, SVF-003.4, SVF- 003.5, and SVF-003.6 delivered in a prime / boost approach. The efficacy of the CCHFV immunogenic compositions was assessed in C57BL / 6 mice, with n=5 mice for each group.

[0140] Plasmids for the DNA constructs were prepared by growing 5L shake flask cultures followed by performing QIAGEN Endofree GIGA plasmid purification, with plasmid amounts depicted in FIG. 10A. DNA compositions comprised SVF-003.1, SVF-003.2, SVF- 003.3, SVF-003.4, SVF-003.5, and SAT-003.6 (SEQ ID NO: 18, 21, 24, 27, 30, or 33). FIG. 10B depicts the prime / boost vaccine delivery strategy for this study. Mice were immunized intramuscularly with 50pg of DNA construct plus adjuvant and subjected to in vivoelectroporation. After two weeks, blood samples were collected from immunized mice and the titers of anti-N antibodies were detected via ELISA and compared to positive and negative controls. As depicted in FIGs. IOC and 10D, immunization with SVF.003.1 -SVF.003.6 DNA compositions resulted in detectable levels of anti-N antibodies at two weeks post immunization.

[0141] Mice received the boost immunization of SVF.003.1 -SVF.003.6 protein at three weeks post initial immunization with the SVF.003.1 -SVF.003.6 DNA constructs. Polypeptide compositions comprised SVF-003.1, SVF-003.2, SVF-003.3, SVF-003.4, SVF- 003.5, and SVF-003.6 (SEQ I® NO: 19, 22, 25, 28, 31, or 34). Mice were sacrificed at five weeks post initial immunization and tissues were collected for further experiments. Titers of anti-N antibodies were detected via ELISA at five weeks post initial immunization and compared to positive and negative controls. As depicted in FIGs. 11 A and 1 IB, anti-N antibody titers were elevated following boost immunization with SVF.003.1-SVF.003.6 protein compositions.

[0142] Purified white blood cells were tested for IFNy production in response to CCHFV antigens by ELISpot. T cell responses were evaluated by collecting splenocytes at five weeks post initial immunization and isolating IFN-y+ and IL-2+ T cells via ELISPOT. Isolated T cells were stimulated with peptide pools comprising the CCHFV M segment and S segment, as well as gp38 protein, Gc protein, and N protein. The proportion of IFN-y+ and IL- 2+ T cells responding to the peptides of the CCHFV peptide pool were assessed. As shown in FIGs. 12A-12H T cells from mice immunized with SVF.003.1 -SVF.003.6 demonstrated an elevated proportion of cells responsive to the N protein pool, and mice immunized with SVF.003.3-SVF.003.6 demonstrated an elevated proportion of cells responsive to the Gc protein pool.

[0143] In at least some of the previously described embodiments, one or more elements used in an embodiment can interchangeably be used in another embodiment unless such a replacement is not technically feasible. It will be appreciated by those skilled in the art that various other omissions, additions and modifications may be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modifications and changes are intended to fall within the scope of the subject matter, as defined by the appended claims.

[0144] With respect to the use of substantially any plural and / or singular terms herein, those having skill in the art can translate from the plural to the singular and / or from the singular to the plural as is appropriate to the context and / or application. The various singular / plural permutations may be expressly set forth herein for sake of clarity.

[0145] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as “open” terms (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases “at least one” and “one or more” to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles “a” or “an” limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases “one or more” or “at least one” and indefinite articles such as “a” or “an” (e.g., “a” and / or “an” should be interpreted to mean “at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of “two recitations,” without other modifiers, means at least, two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to “at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill m the art would understand the convention (e.g., “a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). In those instances where a convention analogous to “at least one of A, B, or C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “ a system having at least one of A, B, or C” would include but not belimited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and / or phrase presenting two or more alternative terms, whether in the description or claims, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase “A or B” will be understood to include the possibilities of “A” or “B” or “A and B.”

[0146] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0147] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1 , 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1 , 2, 3, 4, or 5 articles, and so forth.

[0148] All references cited herein, including but not limited to published and unpublished applications, patents, and literature references, are incorporated herein by reference in their entirety and are hereby made a part of this specification. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material.

[0149] While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects andembodiments disclosed herein are for purposes of illustration and are not intended to be limiting, with the true scope and spirit being indicated by the following claims.REFERENCES1. Duh D, Nichol ST, Khristova ML, Saksida A, Hafner-Bratkovic I, Petrovec M, Dedushaj I, Ahmeti S, Avsic-Zupanc T. The complete genome sequence of a Crimean-Congo hemorrhagic fever virus isolated from an endemic region in Kosovo. Virol J. 2008 Jan 15;5:7. doi: 10.1186 / 1743-422X-5-7.2. Hawman DW’, Ahlen G, Appelberg S, Meade-White K, Hanley P, Scott D, Monteil D, DevignotD, Weber F, Feldmann H, Sallberg M, Mirazhni A. 2021 A DNA-based vaccine protects against Crimean-Congo hemorrhagic fever virus disease in a Cynomolgus macaque model. Nat Microbiol. 2021 Feb;6(2): 187-195. doi: 10.1038 / s41564-020-00815-6.3. Appelberg S, John L, Pardi N, Vegvari A, Bereczky S, Ahlen G, Monteil V, Abdurahman S, Mika el off F, Beatti M, Tam ¥, Sallberg M, Neogi U, Weissman D, Mirazimi A Nucleoside-modified mRNA vaccines protect 1FNAR " ''mice against Crimean Congo hemorrhagic fever virus infection J Virology, 20214. 6. Hawman DW, Meade-White K, Leventhal S, Appelberg S, Ahlen G, Nikoyan N, Clancy C, Smith B, Hanley P, Lovaglio J, Mirazimi A*, Sallberg M*, Feldmann H*. 2022. A two dose DNA vaccine regimen expressing the Crimean-Congo hemorrhagic fever virus nucleoprotein and glycoproteins provides robust protection in a macaque model. Molecular Therapy Molecular Therapy 2022 Oct 3: SI 525- 0016(22)00605-0, doi: 10.1016 / j.ymthe.2022.09.016. * equal contribution5. Golden JW, Shoemaker CJ, Lindquist ME, Zeng X, Daye SP, Williams JA, Liu J, Coffin KM, Olschner S, Flusin O, Altamura LA, Kuehl KA, Fitzpatrick CJ, Schmaljohn CS, Garrison AR. GP38-targetmg monoclonal antibodies protect adult mice against lethal Crimean-Congo hemorrhagic fever virus infection. Sei Adv. 2019 Jul 10;5(7):eaaw9535. doi: 10.1126 / sciadv.aaw9535. eCollection 2019 Jul.6. Suschak JJ, Golden JW, Fitzpatrick CJ, Shoemaker CJ, Badger CV, Schmaljohn CS, Garrison AR. A CCHFV DNA vaccine protects against heterologous challenge and establishes GP38 as immunorelevant in mice. NPJ Vaccines. 2021 Mar 2,6(1):31. doi: 10.1038 / s41541 -021-00293-9.7. MaraveliaP, Frelin L, Ni Y, Noelia Caro Perez1, Ahlen G, Jagya N, Verch G, Verhoye L, Pater L, Johansson M, Pasetto A, Meuleman P, Urban S, and Sallberg M. 2021 . Blocking entry of hepatitis B and D viruses to hepatocytes as a novel immunotherapy for treating chronic infections. Journal of Infectious Diseases, 2021 Jan 4,223(1): 128-138. doi:" 10.1093 / infdis / jiaa036.8. Borm R, Maravelia P, Ahlen G, Ciesek S, Perez NG, Pasetto A, Urban S, van Houtte F, Verhoye L, Wedemeyer H, Johansson M, Frelin L, Sallberg M, and MeulemanP. 2022. Novel prime-boost immune-basedd therapy inhibiting both hepatitis B and D virus infections. Gut DOI: 10.1136 / gutjnl-2022-327216.9. Appelberg S, Ahlen G, Yan J, Nikouyan N, Weber S, Larsson O, Hoglund U, Aleman S, Weber F, Perlhamre E, Apro J, Gidlund EK, Tnvesson O, Salati S,Cadossi M, Tegel H, Hober S, Frelin L, Mirazimi A, and Sallberg M. 2022. A universal SARS-CoV DNA vaccine inducing highly crossreactive neutralizing antibodies and T cells. EMBO Molecular MedicineDOI: 10.15252 / emmm.202215821.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A nucleic acid comprising at least one nucleic acid sequence encoding a CCHFV polypeptide and at least one nucleic acid sequence encoding a P2A autocatalytic polypeptide cleavage site.

2. The nucleic acid of claim 1, wherein the nucleic acid sequences encoding a CCHFV polypeptide comprise a nucleic acid sequence encoding an S-segment polypeptide NP (H), and a nucleic acid sequence encoding an M-segment polypeptide GPS 8 (H).

3. The nucleic acid of any one of claims 1 or 2, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 21, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

4. The nucleic acid of claim 1, wherein the nucleic acid sequences encoding a CCHFV polypeptide comprise a nucleic acid sequence encoding an S-segment polypeptide NP (H), and a nucleic acid sequence encoding an M-segment polypeptide Gc ecto (H).

5. The nucleic acid of any one of claims 1-4, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 27, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

6. The nucleic acid of claim 2, further comprising a nucleic acid sequence encoding an Ig leader sequence.

7. The nucleic acid of any one of claims 1-6, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequenceidentity to SEQ ID NO: 18, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

8. The nucleic acid of claim 4, further comprising a nucleic acid sequence encoding an Ig leader sequence.

9. The nucleic acid of any one of claims 1-8, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 24, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

10. The nucleic acid of claim 4, wherein the nucleic acid sequence encoding a CCHFV polypeptide further comprises a nucleic acid sequence encoding an M-segment polypeptide GP38 (H).11 . The nucleic acid of any one of claims 1-10, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 33, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

12. The nucleic acid of claim 10, further comprising a nucleic acid sequence encoding an Ig leader sequence.

13. The nucleic acid of any one of claims 1-12, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 30, wherein a polypeptide encoded by said nucleic acid comprises an epitope or antigen of CCHFV.

14. A polypeptide comprising at least one CCHV polypeptide sequence and at least one P2A autocatalytic polypeptide cleavage site.

15. The polypeptide of claim 14, wherein the CCHFV polypeptide sequences comprise an S-segment NP (H) polypeptide sequence, and an M-segment GP38 (H) polypeptide sequence.

16. The polypeptide of any one of claims 14 or 15, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 22, wherein the polypeptide comprises an epitope or antigen of CCHFV.

17. The polypeptide of claim 14, wherein the CCHFV polypeptide sequences comprise a S-segment NP (H) polypeptide sequence, and a Gc ecto (H) polypeptide sequence.

18. The polypeptide of any one of claims 14 -17, wherein the polypeptide has at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 28, wherein the polypeptide comprises an epitope or antigen of CCHFV.

19. The polypeptide of claim 15, further comprising an Ig leader polypeptide sequence.

20. The polypeptide of claim 14-19, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 19, wherein the polypeptide comprises an epitope or antigen of CCHFV.

21. The polypeptide of claim 17, further comprising an Ig leader polypeptide sequence.

22. The polypeptide of claim 14-21 , wherein the polypeptide has at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 25, wherein the polypeptide comprises an epitope or antigen of CCHFV.

23. The polypeptide of claim 17, wherein the CCHFV polypeptide sequences further comprise an M-segment GP38 (H) polypeptide sequence.

24. The polypeptide of claim 14-23, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 34, wherein the polypeptide comprises an epitope or antigen of CCHFV.

25. The polypeptide of claim 23, further comprising an Ig leader polypeptide sequence.

26. The polypeptide of claim 14-25, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 31, wherein the polypeptide comprises an epitope or antigen of CCHFV.

27. The nucleic acid of any one of claims 1-13 for use in a medicament, such as for the prevention, treatment or inhibition of CCHFV in a subject, preferably a human.

28. The polypeptide of any one of claims 14-26 for use in a medicament, such as for the prevention, treatment or inhibition of CCHFV in a subject, preferably a human.

29. The polypeptide of any one of claims 14-26 or 28, wherein the polypeptide is recombman tly expressed.

30. The polypeptide of any one of claims 14-26 or 28-29, wherein the polypeptide is recombmantly expressed in a mammalian, bacterial, yeast, insect, or cell-free system.

31. An immunogenic composition or product combination comprising:(a) a nucleic acid comprising at least one nucleic acid sequence encoding a CCHFV polypeptide; and(b) a polypeptide comprising at least one CCHFV polypeptide.

32. The immunogenic composition or product combination of claim 31, wherein the at least one nucleic acid sequence encoding a CCHFV polypeptide comprises: i) a nucleic acid sequence encoding an S-segment NP (H) polypeptide; ii) a nucleic acid sequence encoding an M-segment GP38 (H) polypeptide; iii) a nucleic acid sequence encoding an M-segment Gc ecto (H) polypeptide; iv) a nucleic acid sequence encoding an Ig leader polypeptide; v) a nucleic acid sequence encoding a P2A autocatalytic polypeptide cleavage site; or any combination thereof.

33. The immunogenic composition or product combination of claim 31 or 32, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 18, 21, 24, 27, 30, or 33, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV, which is optionally used in a medicament, such as for the prevention, treatment, or inhibition of CCHFV in a subject, such as a mammal, preferably a human.

34. The immunogenic composition or product combination of any one of claims 31-33, wherein the nucleic acid is codon optimized for expression in a human.

35. The immunogenic composition or product combination of any one of claims 31-34, wherein the at least one CCHFV polypeptide comprises: i) an S-segment NP (H) polypeptide sequence, ii) an M-segment GP38 (H) polypeptide sequence; iii) an M-segment Gc_ecto (H) polypeptide sequence, iv) an Ig leader polypeptide sequence; v) a P2A autocatalytic polypeptide cleavage site sequence; or any combination thereof.

36. The immunogenic composition or product combination of any one of claims 31-35, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99%, or 100% homology or sequence identity to SEQ ID NO: 19, 22, 25, 28, 31 , or 34, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV, which is optionally used in a medicament, such as for the prevention, treatment, or inhibition of CCHFV in a subject, such as a mammal, preferably a human.

37. The immunogenic composition or product combination of any one of claims 31-36, wherein the polypeptide is recombinantly expressed.

38. The immunogenic composition or product combination of any one of claims 31-37, wherein the polypeptide is recombinantly expressed in a mammalian, bacterial, yeast, insect, or cell-free system.

39. The immunogenic composition or product combination of any one of claims 31-38, further comprising an adjuvant,40. The immunogenic composition or product combination of claim 39, wherein the adjuvant is alum.41 . The immunogenic composition or product combination of any one of claims 31-40, wherein the nucleic acid is provided in a recombinant vector.

42. A method of generating an immune response in a subject using the immunogenic composition or product combination set forth in any one of claims 31 -41 , comprising: a) administering to the subject at least, one prime dose comprising the nucleic acid, and b) administering to the subject, at least one boost dose comprising the polypeptide.

43. The method of claim 42, wherein the at least one boost dose further comprises an adjuvant.

44. The method of claim 43, wherein the adjuvant is alum.

45. The method of any one of claims 42-44, wherein the at least one boost dose is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, or 48 days or weeks after the at least one prime dose is administered or within a range of time defined by any two of the aforementioned time points e.g., within 1-48 days or 1-48 weeks.

46. The method of any one of claims 42-45, wherein the administration is provided enteraily, orally, intranasally, parenterally, subcutaneously, intramuscularly, intradermally, or intravenously or any combination thereof, and optionally with in vivo electroporation.

47. The method of any one of claims 42-46, wherein the administration is performed in conjunction with an antiviral therapy.

48. The method of claim 47, wherein the antiviral therapy comprises administration of chloroquine, hydroxychloroquine, favipiravir, favilavir, remdesivir, tocilizumab, galidesivir, sarilumab, lopinavir, ritonavir, darunavir, ribavirin, interferon-a, pegylated interferon-a, or interferon alfa~2b, or any combination thereof.

49. An immunogenic composition or product combination for use in the treatment or inhibition of CCHFV, comprising:(a) a nucleic acid comprising at least one nucleic acid sequence encoding a CCHFV polypeptide; and(b) a polypeptide comprising at least one CCHFV polypeptide.

50. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of claim 49, wherein the at least one nucleic acid sequence encoding a CCHFV polypeptide comprises: i) a nucleic acid sequence encoding an S -segment NP (H) polypeptide ; ii) a nucleic acid sequence encoding an M-segment GP38 (H) polypeptide ; iii) a nucleic acid sequence encoding an M-segment Ge ecto (H) polypeptide ;iv) a nucleic acid sequence encoding an Ig leader polypeptide ; v) a nucleic acid sequence encoding a P2A autocatalytic polypeptide cleavage site; or any combination thereof.

51. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of claim 49 or 50, wherein the nucleic acid has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 18, 21, 24, 27, 30, or 33, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV.

52. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of claim 49 or 51, wherein the nucleic acid is codon optimized for expression in a human.

53. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of claims 49-52, wherein the at least one CCHFV polypeptide comprises: i) an S-segmentNP (H) polypeptide sequence; li) an M-segment GP38 (H) polypeptide sequence; iii) an M-segment Gc_ecto (H) polypeptide sequence; iv) an Ig leader polypeptide sequence; v) a P2A autocatalytic polypeptide cleavage site sequence; or any combination thereof.

54. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of claims 49-53, wherein the polypeptide has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or sequence identity to SEQ ID NO: 19, 22, 25, 28, 31, or 34, wherein the polypeptide comprises an epitope or antigen of CCHFV and, wherein the nucleic acid encodes a polypeptide that comprises an epitope or antigen of CCHFV.

55. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of claims 49-54, wherein the polypeptide is recombinantly expressed.

56. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of claims 49-55, wherein the polypeptide is recombinantly expressed in a mammalian, bacterial, yeast, insect, or cell-free system.

57. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of claims 49-56, further comprising an adjuvant.

58. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of claim 57, wherein the adjuvant is alum.

59. The immunogenic composition or product combination for use in the treatment or inhibition of CCHFV of any one of claims 49-58, wherein the nucleic acid is provided in a recombinant vector.

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